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Literature summary extracted from

  • Zhang, Y.; Guo, J.; Zhang, N.; Yuan, W.; Lin, Z.; Huang, W.
    Characterization and analysis of a novel diguanylate cyclase PA0847 from Pseudomonas aeruginosa PAO1 (2019), Infect. Drug Resist., 12, 655-665 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.65 gene PA0847, recombinant expression of wild-type and mutant enzymes in PSeusomonas aeruginosa strain PAO1, complementation of PA0847 is applied by amplification of the full-length gene with its native promoter Pseudomonas aeruginosa

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.65 D609A site-directed mutagenesis, inactive mutant Pseudomonas aeruginosa
2.7.7.65 D626A site-directed mutagenesis, inactive mutant Pseudomonas aeruginosa
2.7.7.65 D652A site-directed mutagenesis, inactive mutant Pseudomonas aeruginosa
2.7.7.65 K614A site-directed mutagenesis, inactive mutant Pseudomonas aeruginosa
2.7.7.65 K722A site-directed mutagenesis, inactive mutant Pseudomonas aeruginosa
2.7.7.65 additional information construction and expression of the PA0847 transposon mutants DELTAPA0847-1 and DELTAPA0847-2 in Pseudomonas aeruginosa strain PAO1, comparison to wild-type, phenotypes, overview. Generation of an enzyme deletion mutant, DELTAPA0847 Pseudomonas aeruginosa
2.7.7.65 R595A site-directed mutagenesis, the mutant shows similar activity compared to wild-type enzyme Pseudomonas aeruginosa
2.7.7.65 R641A site-directed mutagenesis, inactive mutant Pseudomonas aeruginosa
2.7.7.65 R641A/R644A site-directed mutagenesis, the mutant shows similar activity compared to wild-type enzyme Pseudomonas aeruginosa
2.7.7.65 R672A site-directed mutagenesis, the mutant shows increased activity compared to wild-type enzyme Pseudomonas aeruginosa

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.7.65 c-di-GMP weak product feedback inhibition is found in PA0847 GGDEF domain suggesting that allosteric regulation from the sensory domain could play a dominant role in the control of intracellular c-di-GMP synthesis Pseudomonas aeruginosa
2.7.7.65 additional information enzyme PA0847 demonstrates no significant product inhibition, though the key residues of two I-sites for c-di-GMP binding are conserved in its GGDEF domain. ENzyme PA0847 contains the canonical product inhibition site (primary I site) of the RxxD motif, which is located upstream of the conserved GGDEF motif. It also has a secondary inhibition site which is conserved in GGDEF domains that can undergo feedback inhibition. These two sites may form a pocket for c-di-GMP binding Pseudomonas aeruginosa

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.7.7.65 membrane domain analysis of PA0847 shows a unique molecular architecture with an N-terminal periplasmic sensory domain and a C-terminal intracellular GGDEF domain Pseudomonas aeruginosa 16020
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.7.65 Mg2+ required Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.7.65 2 GTP Pseudomonas aeruginosa
-
2 diphosphate + cyclic di-3',5'-guanylate
-
?
2.7.7.65 2 GTP Pseudomonas aeruginosa ATCC 15692
-
2 diphosphate + cyclic di-3',5'-guanylate
-
?
2.7.7.65 2 GTP Pseudomonas aeruginosa 1C
-
2 diphosphate + cyclic di-3',5'-guanylate
-
?
2.7.7.65 2 GTP Pseudomonas aeruginosa PRS 101
-
2 diphosphate + cyclic di-3',5'-guanylate
-
?
2.7.7.65 2 GTP Pseudomonas aeruginosa DSM 22644
-
2 diphosphate + cyclic di-3',5'-guanylate
-
?
2.7.7.65 2 GTP Pseudomonas aeruginosa CIP 104116
-
2 diphosphate + cyclic di-3',5'-guanylate
-
?
2.7.7.65 2 GTP Pseudomonas aeruginosa LMG 12228
-
2 diphosphate + cyclic di-3',5'-guanylate
-
?
2.7.7.65 2 GTP Pseudomonas aeruginosa JCM 14847
-
2 diphosphate + cyclic di-3',5'-guanylate
-
?

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.65 Pseudomonas aeruginosa Q9I594
-
-
2.7.7.65 Pseudomonas aeruginosa 1C Q9I594
-
-
2.7.7.65 Pseudomonas aeruginosa ATCC 15692 Q9I594
-
-
2.7.7.65 Pseudomonas aeruginosa CIP 104116 Q9I594
-
-
2.7.7.65 Pseudomonas aeruginosa DSM 22644 Q9I594
-
-
2.7.7.65 Pseudomonas aeruginosa JCM 14847 Q9I594
-
-
2.7.7.65 Pseudomonas aeruginosa LMG 12228 Q9I594
-
-
2.7.7.65 Pseudomonas aeruginosa PRS 101 Q9I594
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.65 2 GTP
-
Pseudomonas aeruginosa 2 diphosphate + cyclic di-3',5'-guanylate
-
?
2.7.7.65 2 GTP
-
Pseudomonas aeruginosa ATCC 15692 2 diphosphate + cyclic di-3',5'-guanylate
-
?
2.7.7.65 2 GTP
-
Pseudomonas aeruginosa 1C 2 diphosphate + cyclic di-3',5'-guanylate
-
?
2.7.7.65 2 GTP
-
Pseudomonas aeruginosa PRS 101 2 diphosphate + cyclic di-3',5'-guanylate
-
?
2.7.7.65 2 GTP
-
Pseudomonas aeruginosa DSM 22644 2 diphosphate + cyclic di-3',5'-guanylate
-
?
2.7.7.65 2 GTP
-
Pseudomonas aeruginosa CIP 104116 2 diphosphate + cyclic di-3',5'-guanylate
-
?
2.7.7.65 2 GTP
-
Pseudomonas aeruginosa LMG 12228 2 diphosphate + cyclic di-3',5'-guanylate
-
?
2.7.7.65 2 GTP
-
Pseudomonas aeruginosa JCM 14847 2 diphosphate + cyclic di-3',5'-guanylate
-
?

Subunits

EC Number Subunits Comment Organism
2.7.7.65 dimer domain analysis of PA0847 shows a unique molecular architecture with an N-terminal periplasmic sensory domain and a C-terminal intracellular GGDEF domain. Structural model of PA0847 GGDEF dimer, overview Pseudomonas aeruginosa

Synonyms

EC Number Synonyms Comment Organism
2.7.7.65 DGC
-
Pseudomonas aeruginosa
2.7.7.65 GGDEF domain-containing protein
-
Pseudomonas aeruginosa
2.7.7.65 PA0847
-
Pseudomonas aeruginosa

Temperature Optimum [┬░C]

EC Number Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
2.7.7.65 37
-
assay at Pseudomonas aeruginosa

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.7.65 7.6
-
assay at Pseudomonas aeruginosa

General Information

EC Number General Information Comment Organism
2.7.7.65 malfunction weak product feedback inhibition is found in PA0847 GGDEF domain suggesting that allosteric regulation from the sensory domain could play a dominant role in the control of intracellular c-di-GMP synthesis Pseudomonas aeruginosa
2.7.7.65 additional information the diguanylate cyclase activity of PA0847 depends on the neighboring PAS domain. A local structural change imposed by an adjacent tyrosine residue is identified, which indicates the structural and functional diversities of the GGDEF family proteins. Signaling mechanism of the unique c-di-GMP metabolizing protein PA0847, overview. Three-dimensional homology model of PA0847 GGDEF domain (residues 576-736) using DGC DgcZ (PDB ID 4H54) as a structure template. Seven critical residues, D609, K614, N617, D626, D652, E653, and K722, are involved in GTP binding Pseudomonas aeruginosa
2.7.7.65 physiological function the enzyme synthesizes the central signaling molecule, cyclic diguanylate (c-di-GMP), which is found to regulate various bacterial phenotypes, especially those involved in pathogen infection and drug resistance. Enzyme PA0847 is an active diguanylate cyclase (DGC). Roles of PA0847 in the regulation of bacterial motility and biofilm formation. Enzyme PA0847 is involved in response to a variety of environmental nutrients and factors, suggesting it might serve as a broad-range environmental sensor on the membrane of Pseudomonas aeruginosa. Gene of PA0847 affects biofilm formation and motility phenotypes in Pseudomonas aeruginosa Pseudomonas aeruginosa